Mouse M-CSF/CSF1 enzyme-linked immunoassay kit

CAT: EM0056 Datasheet
Specification 96 Test
Sensitivity 0.31 pg/ml (10 μl)
Standard Curve Range 6.86~5000 pg/ml
Standard Curve Gradient 7 Points/3 Folds
Number of Incubations 2
Detectable sample Liquid phase sample of soluble substances. For example: serum, plasma, cell culture supernatant, tissue grinding liquid, etc.
Sample Volume 10 μl
Type Fully Ready-to-Use
Operation Duration 120min
pg/ml O.D. Average Corrected
0.00 0.0153 0.0169 0.0161
6.86 0.0487 0.0427 0.0457 0.0296
20.58 0.1196 0.1191 0.1194 0.1033
61.73 0.3429 0.3280 0.3355 0.3194
185.19 0.9188 0.8179 0.8684 0.8523
555.56 2.1906 2.2550 2.2228 2.2067
1666.67 3.7043 3.3983 3.5513 3.5352
5000.00 4.1075 4.2155 4.1615 4.1454

Precision

Intra-assay Precision Inter-assay Precision
Sample Number S1 S2 S3 S1 S2 S3
22 22 22 6 6 6
Average(pg/ml) 95.79 470.48 1171.32 97.6 458.2 1089.7
Standard Deviation 3.24 24.57 71.90 4.3 23.9 60.5
Coefficient of Variation(%) 3.4 5.2 6.1 4.4 5.2 5.6

Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.

Inter-assay Precision (Precision between assays) Three samples of known concentration were tested six times on one plate to assess intra-assay precision.

Spike Recovery

The spike recovery was evaluated by spiking 3 levels of mouse M-CSF/CSF1 into health mouse serum sample. The un-spiked serum was used as blank in this experiment.
The recovery ranged from 83% to 115% with an overall mean recovery of 96%.

Sample Values

Sample Matrix Sample Evaluated Range (pg/ml) Detectable (%) Mean of Detectable (pg/ml)
Serum 30 1419.67-3710.99 100 2145.38

Serum/Plasma – Thirty samples from apparently healthy mice were evaluated for the presence of M-CSF/CSF1 in this assay. No medical histories were available for the donors. n.d. = non-detectable. Samples measured below the sensitivity are considered to be non-detectable.

Background: M-CSF/CSF1

M-CSF, also known as CSF-1, is a four-alpha-helical-bundle cytokine that is the primary regulator of macrophage survival, proliferation and differentiation. M-CSF is also essential for the survival and proliferation of osteoclast progenitors. M-CSF also primes and enhances macrophage killing of tumor cells and microorganisms, regulates the release of cytokines and other inflammatory modulators from macrophages, and stimulates pinocytosis. M-CSF increases during pregnancy to support implantation and growth of the decidua and placenta. Sources of M-CSF include fibroblasts, activated macrophages, endometrial secretory epithelium, bone marrow stromal cells and activated endothelial cells. The M-CSF receptor (c-fms) transduces its pleotropic effects and mediates its endocytosis. M-CSF mRNAs of various sizes occur. Full length human M-CSF transcripts encode a 522 amino acid (aa) type I transmembrane (TM) protein with a 464 aa extracellular region, a 21 aa TM domain, and a 37 aa cytoplasmic tail that forms a 140 kDa covalent dimer. Differential processing produces two proteolytically cleaved, secreted dimers. One is an N- and O- glycosylated 86 kDa dimer, while the other is modified by both glycosylation and chondroitin-sulfate proteoglycan (PG) to generate a 200 kDa subunit. Although PG-modified M-CSF can circulate, it may be immobilized by attachment to type V collagen. Shorter transcripts encode M-CSF that lack cleavage and PG sites and produce an N-glycosylated 68 kDa TM dimer and a slowly produced 44 kDa secreted dimer. Although forms may vary in activity and half-life, all contain the N-terminal 150 aa portion that is necessary and sufficient for interaction with the M-CSF receptor. The first 223 aa of mature human M-CSF shares 88%, 86%, 81% and 74% aa identity with corresponding regions of dog, cow, mouse and rat M-CSF, respectively. Human M-CSF is active in the mouse, but mouse M-CSF is reported to be species-specific.

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